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Strategic single point mutation yields a solvent- and salt-stable transaminase from Virgibacillus sp. in soluble form

Guidi, Benedetta; Planchestainer, Matteo; Contente, Martina Letizia; Laurenzi, Tommaso; Eberini, Ivano; Gourlay, Louise J.; Romano, Diego; Paradisi, Francesca; Molinari, Francesco

Authors

Benedetta Guidi

Matteo Planchestainer

Martina Letizia Contente

Tommaso Laurenzi

Ivano Eberini

Louise J. Gourlay

Diego Romano

Francesca Paradisi

Francesco Molinari



Abstract

A new transaminase (VbTA) was identified from the genome of the halotolerant marine bacterium Virgibacillus 21D and its expression resulted in the production of an insoluble protein in E. coli. After a single mutation identified through sequence homology analysis, the mutant VbTA T16F was successfully expressed as a recombinant protein and characterised. VbTA T16F showed high stability towards polar organic solvents and salt exposure, accepting mainly hydrophobic aromatic amine and carbonyl substrates. The 2.0 Å resolution crystal structure of VbTA T16F is reported, and together with computational calculations, revealed that this mutation is crucial for correct dimerisation and thus correct folding, leading to soluble protein expression.

Journal Article Type Article
Publication Date Dec 1, 2018
Journal Scientific Reports
Print ISSN 2045-2322
Electronic ISSN 2045-2322
Publisher Nature Research
Peer Reviewed Peer Reviewed
Volume 8
Issue 1
Article Number 16441
APA6 Citation Guidi, B., Planchestainer, M., Contente, M. L., Laurenzi, T., Eberini, I., Gourlay, L. J., …Molinari, F. (2018). Strategic single point mutation yields a solvent- and salt-stable transaminase from Virgibacillus sp. in soluble form. https://doi.org/10.1038/s41598-018-34434-3
DOI https://doi.org/10.1038/s41598-018-34434-3
Publisher URL https://www.nature.com/articles/s41598-018-34434-3

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